Expression Techniques (expression + techniques)

Distribution by Scientific Domains


Selected Abstracts


Promoter analysis of ventricular myosin heavy chain (vmhc) in zebrafish embryos

DEVELOPMENTAL DYNAMICS, Issue 7 2009
Daqing Jin
Abstract In zebrafish, ventricular myosin heavy chain (vmhc) gene is initially expressed at the anterior lateral mesoderm and thereafter its expression is restricted to the cardiac ventricle. The transcriptional control mechanisms in regulating chamber-specific expression of myosin heavy chains are not well defined. We isolated and analyzed zebrafish vmhc upstream region to examine the spatial and temporal regulation of vmhc using transgenic and transient expression techniques. Promoter deletion analyses defined a basal promoter region sufficient to drive vmhc expression in the ventricle and an upstream fragment necessary for repressing ectopic vmhc expression in the atrium. The transcriptional mechanism that prevents vmhc expression in the atrium is mediated through Nkx2.5 binding elements (NKE). We have further discovered that paired-related homeobox transcriptional factor 2 (Prx2/S8)-like binding elements are required for promoting vmhc expression, and Prrx1b, a Prx-related homeobox protein, participates in the regulation of vmhc expression with other transcriptional factors. Developmental Dynamics 238:1760,1767, 2009. © 2009 Wiley-Liss, Inc. [source]


DOE genomics: Applications to in situ subsurface bioremediation

REMEDIATION, Issue 1 2006
Robert T. AndersonArticle first published online: 22 DEC 200
Microbial communities can greatly affect the mobility and fate of subsurface contaminants, yet relatively little is known about the functioning of microorganisms in subsurface environments. Major advances in DNA sequencing capability and the advent of genome-enabled studies have produced key insights into how microorganisms adapt to environmental conditions and/or biotransform subsurface contaminants starting from analyses of genome content. These techniques enable the researcher to detect how an organism responds to its environment and, potentially, to devise better methods to promote specific microbial activity in subsurface environments. The U.S. Department of Energy sponsors genome research through the Genomics:GTL program. One of the applications of this research is to better understand and control biological processes influencing the mobility of contaminants of concern to DOE such as metals and radionuclides. Genome and gene expression techniques have led to new insights into the functioning of subsurface microbial communities, but the true potential of these techniques is still to be revealed. As genome-enabled science progresses, techniques for evaluating gene expression patterns of whole communities will advance the understanding and development of optimized in situ bioremediation and more realistic simulations of microbial contaminant biotransformation. © 2006 Wiley Periodicals, Inc.* [source]


Preconditioning of skeletal muscle against contraction-induced damage: the role of adaptations to oxidants in mice

THE JOURNAL OF PHYSIOLOGY, Issue 1 2004
F. McArdle
Adaptations of skeletal muscle following exercise are accompanied by changes in gene expression, which can result in protection against subsequent potentially damaging exercise. One cellular signal activating these adaptations may be an increased production of reactive oxygen and nitrogen species (ROS). The aim of this study was to examine the effect of a short period of non-damaging contractions on the subsequent susceptibility of muscle to contraction-induced damage and to examine the changes in gene expression that occur following the initial contraction protocol. Comparisons with changes in gene expression in cultured myotubes following treatment with a non-damaging concentration of hydrogen peroxide (H2O2) were used to identify redox-sensitive genes whose expression may be modified by the increased ROS production during contractions. Hindlimb muscles of mice were subjected to a preconditioning, non-damaging isometric contraction protocol in vivo. After 4 or 12 h, extensor digitorum longus (EDL) and soleus muscles were removed and subjected to a (normally) damaging contraction protocol in vitro. Muscles were also analysed for changes in gene expression induced by the preconditioning protocol using cDNA expression techniques. In a parallel study, C2C12 myotubes were treated with a non-damaging concentration (100 ,m) of H2O2 and, at 4 and 12 h following treatment, myotubes were treated with a damaging concentration of H2O2 (2 mm). Myotubes were analysed for changes in gene expression at 4 h following treatment with 100 ,m H2O2 alone. Data demonstrate that a prior period of non-damaging contractile activity resulted in significant protection of EDL and soleus muscles against a normally damaging contraction protocol 4 h later. This protection was associated with significant changes in gene expression. Prior treatment of myotubes with a non-damaging concentration of H2O2 also resulted in significant protection against a damaging treatment, 4 and 12 h later. Comparison of changes in gene expression in both studies identified haem oxygenase-1 as the sole gene showing increased expression during adaptation in both instances suggesting that activation of this gene results from the increased ROS production during contractile activity and that it may play a role in protection of muscle cells against subsequent exposure to damaging activity. [source]


Manufacturing antibodies in the plant cell

BIOTECHNOLOGY JOURNAL, Issue 12 2009
Diego Orzáez Dr.
Abstract Plants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently producing mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being overcome. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability into highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production. [source]


Heterologous GPCR Expression: A Bottleneck to Obtaining Crystal Structures

BIOTECHNOLOGY PROGRESS, Issue 3 2007
Emily C. McCusker
G protein-coupled receptors (GPCRs) are an important, medically relevant class of integral membrane proteins. Laboratories throughout all disciplines of science devote time and energy into developing practical methods for the discovery, isolation, and characterization of these proteins. Since the crystal structure of rhodopsin was solved 6 years ago, the race to determine high-resolution structures of more GPCRs has gained momentum. Since certain GPCRs are currently produced at sufficient levels for X-ray crystallography trials, it is speculated that heterologous expression of GPCRs may no longer be a bottleneck in obtaining crystal structures. This Review focuses on the current approaches in heterologous expression of GPCRs and explores the problems associated with obtaining crystal structures from GPCRs expressed in different systems. Although milligram amounts of certain GPCRs are attainable, the majority of GPCRs are still either produced at very low levels or not at all. Developing reliable expression techniques for GPCRs is still a major priority for the structural characterization of GPCRs. [source]